The Atomic Force Microscope (AFM) can image and manipulate biological samples under physiological conditions at nanometer resolution. Despite its very slow operation (sometimes over 30 minutes per image) and the special expertise required to overcome its complexity of use, the AFM has already facilitated significant biomedical discoveries including new insights into membrane proteins and the mechanical properties of structural proteins. The range of biological processes the AFM can image is, however, severely limited by the AFM's current low time resolution, which is due to fundamental limits inherent in the current generation of AFMs based on cantilevers with dimensions of order 100 microns. This new generation of user-friendly (2 hours'training), high speed (2 images/second) AFMs is proposed to be developed based on fundamental innovations in design, particularly by basing the new AFMs on cantilevers with dimensions of order 10 microns to improve time and force resolution for probing molecules and speed in imaging by an order of magnitude or more. The Specific Aims of the project are: i) to develop a high-speed AFM that can image at 512x512 resolution, at 2 images/second;2) to develop a user-friendly AFM in which most of the processes in setting up the AFM and adjusting feedback parameters are automated;3) to develop a new AFM that combines high-speed with automation. It is hoped and expected that these new AFMs can serve as prototypes to guide the development of future commercial AFMs, which can bring the advantages of high-speed and automation to the broader community of biomedical researchers. This new generation of AFMs will be fast and easy to use and have improved tune and force resolution. This will facilitate imaging of biological processes at the molecular level and thus have a broad enabling impact on biomedical research. The long term goal is to transition the AFM from a specialized research tool to a general use medical instrument.